Abstract
Background
The non-structural protein 4 (NSP4) of porcine reproductive and respiratory syndrome virus (PRRSV) possesses 3C-like serine protease (3CLSP) activity, which can influence PRRSV replication, suppress host IFN-β production, induce host cell apoptosis, and play a crucial role in PRRSV detections. Wild or attenuated vaccine strains will produce antibodies against non-structural proteins, while inactivated vaccines will not produce antibodies against non-structural proteins. The Enzyme-Linked Immunosorbnent Assay (ELISA) method for non-structural proteins can distinguish immunity effect of inactivated vaccine from wild strain or attenuated vaccine strains. Antibodies induced by NSP4 can effectively serve as indicators of infections caused by the wild-type virus. In the present study, the NSP4 protein from the PRRSV XH-GD strain (GenBank No. EU624117.1) was cloned, expressed, and used as a coating protein to establish an indirect enzyme-linked immunosorbent assay (ELISA) detection method. The specificity, repeatability, sensitivity, and agreement rates with those of commercial ELISA kits were compared in this study.
Results
The developed NSP4 indirect ELISA method displayed excellent specificity, repeatability, and sensitivity, with an impressive agreement rate of 91.74% with the PRRSV IDEXX ELISA kit.
Conclusion
The indirect ELISA method for PRRSV NSP4 was successfully constructed., Utilizing the PRRSV NSP4s to establish an ELISA antibody detection method was a more conducive for sustained antibody monitoring in pig farms over time Therefore, the establishment of an NSP4 indirect ELISA detection method provides technical support for the detection of PRRSV antibodies. The coincidence rate between this method and commercial kit is high, which lays a foundation for distinguishing inactivated vaccine from attenuated vaccine.